ß-Elemene induces apoptosis and autophagy in colorectal cancer cells through regulating the ROS/AMPK/mTOR pathway.

ß-Elemene is an effective anti-cancer ingredient extracted from the genus Curcuma (Zingiberaceae familiy). In the present study, we demonstrated that ß-elemene inhibited the proliferation of colorectal cancer cells and induced cell cycle arrest in the G2/M phase. In addition, ß-elemene induced nuclear chromatin condensation and cell membrane phosphatidylserine eversion, decreased cell mitochondrial membrane potential, and promoted the cleavage of caspase-3, caspase-9 and PARP proteins, indicating apoptosis in colorectal cancer cells. At the same time, ß-elemene induced autophagy response, and the treated cells showed autophagic vesicle bilayer membrane structure, which was accompanied by up-regulation of the expression of LC3B and SQSTM1. Furthermore, ß-elemene increased ROS levels in colorectal cancer cells, promoted phosphorylation of AMPK protein, and inhibited mTOR protein phosphorylation. In the experiments in vivo, ß-elemene inhibited the tumor size and induced apoptosis and autophagy in nude mice. In summary, ß-elemene inhibited the occurrence and development of colon cancer xenografts in nude mice, and significantly induced apoptosis and autophagy in colorectal cancer cells in vitro. These effects were associated with regulation of the ROS/AMPK/mTOR signaling. We offered a molecular basis for the development of ß-elemene as a promising anti-tumor drug candidate for colorectal cancer.

Minimally invasive treatment of hepatic hemangioma by transcatheter arterial embolization combined with microwave ablation: A case report.

BACKGROUND: Hepatic hemangioma is the most common benign tumor of the liver. However, patients with large hemangiomas that cause compression symptoms or that are at risk of rupture may need further intervention. It is necessary to explore additional minimally invasive and personalized treatment options for hemangiomas. CASE SUMMARY: A 47-year-old woman was diagnosed with a right hepatic hemangioma for more than 10 years. Abdominal contrast-enhanced computed tomography (CT) and contrast-enhanced ultrasound revealed that there was a large hemangioma in the right liver, with a size of approximately 95 mm × 97 mm × 117 mm. Due to the patient's refusal of surgical treatment, hepatic artery embolization was performed in the first stage. After 25 d of liver protection treatment, the liver function indexes decreased to normal levels. Then, ultrasound-guided microwave ablation of the giant hepatic hemangioma was performed. Ten days after the treatment, hepatobiliary ultrasonography showed that the hemangioma of the right liver was smaller than the previous size (the volume was reduced by approximately 30%). Then the patient was discharged from the hospital. One year after discharge, CT showed that the hepatic hemangioma had shrunk by about 80. CONCLUSION: Transcatheter arterial embolization combined with microwave ablation is a safe and effective minimally invasive treatment for hepatic hemangioma.

[Expression of Semaphorin 3A after spinal cord injury].

OBJECTIVE: To investigate expression of Semaphorin 3A in rats after spinal cord injury and explore possible mechanism of inhibiting of axonal regeneration after SCI. METHODS: Forty healthy female SD rats, 8 weeks old, weighing (210.00±9.88) g, were randomly divided into control group(20 rats in group A) and model group(20 rats in group B). In control group, removal of T10 lamina and partial removal of T9 and T11 lamina were performed, and no further operation was performed on spinal cord (pseudo operation). In model group, the total T10 and partial T9, T11 partial lamina were incised and the spinal cord transection was performed to create animal models of spinal cord injury. The rats were perfused and spinal cord tissue obtained at 3, 7, 14, 28 and 42 days after surgery (4 rats in each group at each time point), respectively, and then HE staining was performed. Meanwhile, the expression of Semaphoring 3A was detected in accordance with the protocol of SP kit. RESULTS: After a simple spinal cord transection injury, hemorrhagic necrosis, localized edema, neurodegeneration, necrosis, and cyst formation occurred in the injured area, and glial scar formation occurred in glial cells. Semaphorin 3A expression levels in control group was low in the gray matter area. There was no expression of Semaphorin 3A in the injured area of spinal cord injury in model group 3 days after operation. On the 14th day, the expression of Semaphorin 3A in the injured area of spinal cord injury increased significantly and was at a high level. On the 28th day, the expression of Semaphorin 3A was moderate. On the 42th day, the positive expression of Semaphorin 3A returned to normal level. CONCLUSION: The increased expression of Semaphorin 3A after spinal cord injury may be one of the mechanisms that inhibit axonal regeneration.

Olfactory ensheathing cell transplantation alters the expression of chondroitin sulfate proteoglycans and promotes axonal regeneration after spinal cord injury.

Cell transplantation is a potential treatment for spinal cord injury. Olfactory ensheathing cells (OECs) play an active role in the repair of spinal cord injury as a result of the dual characteristics of astrocytes and Schwann cells. However, the specific mechanisms of repair remain poorly understood. In the present study, a rat model of spinal cord injury was established by transection of T10. OECs were injected into the site, 1 mm from the spinal cord stump. To a certain extent, OEC transplantation restored locomotor function in the hindlimbs of rats with spinal cord injury, but had no effect on the formation or volume of glial scars. In addition, OEC transplantation reduced the immunopositivity of chondroitin sulfate proteoglycans (neural/glial antigen 2 and neurocan) and glial fibrillary acidic protein at the injury site, and increased the immunopositivity of growth-associated protein 43 and neurofilament. These findings suggest that OEC transplantation can regulate the expression of chondroitin sulfate proteoglycans in the spinal cord, inhibit scar formation caused by the excessive proliferation of glial cells, and increase the numbers of regenerated nerve fibers, thus promoting axonal regeneration after spinal cord injury. The study was approved by the Animal Ethics Committee of the Medical College of Xi'an Jiaotong University, China (approval No. 2018-2048) on September 9, 2018.

Adult duodenal intussusception with horizontal adenoma: A rare case report.

BACKGROUND: Adult duodenal intussusception rarely occurs, and the majority of duodenal adenomas are located in the descending part of the duodenum. Therefore, adenomas in the horizontal part of the duodenum presenting as duodenal intussusception in adults are extremely rare. CASE SUMMARY: A 36-year-old man complained of abdominal pain for 13 d. Blood analysis showed anemia. Magnetic resonance cholangiopancreatography and computed tomography revealed a tumor in the horizontal part of the duodenum as the main finding, leading to duodeno-duodenal intussusception. No obvious abnormalities were found on endoscopy or upper gastrointestinal radiography. He was diagnosed with duodenal intussusception secondary to duodenal adenoma. Laparotomy showed duodeno-duodenal intussusception and a tumor in the horizontal part of the duodenum near the ascending part. Postoperative pathology revealed tubular-villous adenoma with low-grade glandular intraepithelial neoplasia (local high-grade intraepithelial neoplasia). He was discharged without complications. CONCLUSION: This case highlights that rational use of computed tomography, magnetic resonance cholangiopancreatography, endoscopy and upper gastrointestinal radiography for preoperative diagnosis and timely surgery is an effective strategy for the treatment of adult duodenal intussusception with duodenal masses.

Urinary Bile Acid Profile of Newborns Born by Cesarean Section Is Characterized by Oxidative Metabolism of Primary Bile Acids: Limited Roles of Fetal-Specific CYP3A7 in Cholate Oxidations.

This work aims to investigate how the bile acid metabolism of newborns differs from that of adults along the axis of primary, secondary, and tertiary bile acids (BAs). The total unconjugated BA profiles were quantitatively determined by enzyme digestion techniques in urine of 21 newborns born by cesarean section, 29 healthy parturient women, 30 healthy males, and 28 healthy nonpregnant females. As expected, because of a lack of developed gut microbiota, newborns exhibited poor metabolism of secondary BAs. Accordingly, the tertiary BAs contributed limitedly to the urinary excretion of BAs in newborns despite their tertiary-to-secondary ratios significantly increasing. As a result, the primary BAs of newborns underwent extensive oxidative metabolism, resulting in elevated urinary levels of some fetal-specific BAs, including 3-dehydroCA, 3ß,7α,12α-trihydroxy-5ß-cholan-24-oic acid, 3α,12-oxo-hydroxy-5ß-cholan-24-oic acid, and nine tetrahydroxy-cholan-24-oic acids (Tetra-BAs). Parturient women had significantly elevated urinary levels of tertiary BAs and fetal-specific BAs compared with female control, indicating that they may be excreted into amniotic fluid for maternal disposition. An in vitro metabolism assay in infant liver microsomes showed that four Tetra-BAs and 3-dehydroCA were hydroxylated metabolites of cholate, glycocholate, and particularly taurocholate. However, the recombinant cytochrome P450 enzyme assay found that the fetal-specific CYP3A7 did not contribute to these oxidation metabolisms as much as expected compared with CYP3A4. In conclusion, newborns show a BA metabolism pattern predominated by primary BA oxidations due to immaturity of secondary BA metabolism. Translational studies following this finding may bring new ideas and strategies for both pediatric pharmacology and diagnosis and treatment of perinatal cholestasis-associated diseases. SIGNIFICANCE STATEMENT: The prenatal BA disposition is different from adults because of a lack of gut microbiota. However, how the BA metabolism of newborns differs from that of adults along the axis of primary, secondary, and tertiary BAs remains poorly defined. This work demonstrated that the urinary BA profiles of newborns born by cesarean section are characterized by oxidative metabolism of primary BAs, in which the fetal-specific CYP3A7 plays a limited role in the downstream oxidation metabolism of cholate.

Xanthatin mediates G2/M cell cycle arrest, autophagy and apoptosis via ROS/XIAP signaling in human colon cancer cells.

Xanthatin is a natural plant bicyclic sesquiterpene lactone extracted from Xanthium plants (Asteraceae). In the present study, we demonstrated for the first time that Xanthatin inhibited cell proliferation and mediated G2/M phase arrest in human colon cancer cells. Xanthatin also activated caspase and mediated apoptosis in these cells. Concomitantly, Xanthatin triggered cell autophagic response. We found down-regulation of X-linked inhibitor of apoptosis protein (XIAP) contribute to the induction of apoptosis and autophagy. Moreover, reactive oxygen species (ROS) production was triggered upon exposure to Xanthatin in colon cancer cells. ROS inhibitor N-acetylcysteine (NAC) significantly reversed Xanthatin-mediated XIAP down-regulation, G2/M phase arrest, apoptosis and autophagosome accumulation. In summary, our findings demonstrated that Xanthatin caused G2/M phase arrest and mediated apoptosis and autophagy through ROS/XIAP in human colon cancer cells. We provided molecular bases for developing Xanthatin as a promising antitumor candidate for colon cancer therapy. AbbreviationsROSreactive oxygen speciesDMSOdimethyl sulfoxide5-FU5-Fluorouracil3-MA3-MethyladenineDCFH-DA2'7'-dichlorfluorescein-diacetateNACN-acetylcysteineXIAPX-linked inhibitor of apoptosis protein.

GINS2 promotes cell proliferation and inhibits cell apoptosis in thyroid cancer by regulating CITED2 and LOXL2.

To explore the mechanisms of GINS2 on cell proliferation and apoptosis in thyroid cancer (TC) cells. Expressions of GINS2 were inhibited in K1 and SW579 cells using gene interference technology. The abilities of proliferation and apoptosis, and cell cycle were determined by MTT assay and flow cytometric assay. The downstream molecules of GINS2 were searched by microarray and bioinformatics and validated by qRT-PCR and western blotting. In the in vivo study, the tumor growth was compared and the whole-body fluorescent imaging was analyzed. After GINS2 was interfered, cell proliferation was significantly inhibited (P < 0.01) and apoptosis rate increased (P < 0.01) in both K1 and SW579 cells. Cell cycle changed significantly in K1 cells, but not in SW579 cells. With bioinformatics upstream analysis, TGF-ß1 was found as the most significantly upstream regulator. Expressions of TGF-ß1 and its downstream target molecules CITED2 and LOXL2 were validated and found downregulated significantly in mRNA and protein levels (P < 0.05). The results of the nude mouse xenograft assay suggested that the volume and weight of tumor in ones infected with shGINS2 were statistically smaller than controls (P < 0.05). GINS2 plays an important role in cell proliferation and apoptosis of thyroid cancer by regulating the expressions of CITED2 and LOXL2, which may be a potential biomarker for diagnosis or prognosis and a drug target for therapy.

[Preparation and release mechanism of tectorigenin intragastric floating sustained-release tablets].

To investigate the preparation technology and release mechanism of tectorigenin intragastric floating sustained-release tablets. The tablet was produced by wet granulation compression method, with hydroxypropyl methyl cellulose (HPMCK15M), cross-linked polyvinyl pyrrolidone (PVPP), octadecanol and sodium bicarbonate as excipient. The prescriptions were screened and optimized by orthogonal experimental design with in vitro floating capacity and drug release characteristics as the evaluation indexes. The optimization results were as follows: tectorigenin 33.3%, HPMCK15M 16.7%, PVPP 20.0%, octadecanol 13.3%, sodium bicarbonate 5%, and starch gel 10.7%. The prepared tablet can be floated within 10 s in the artificial gastric juice, lasting for 12 h in vitro, with a cumulative release rate of 70% in 10 h. The analysis of Rritger-Peppas equation showed that the sustained-release tablet had two advantages of both drug diffusion and skeleton dissolution. The tablet had good appearance and compressibility, as well as favorable floating capacity and drug release characteristics.

Image-guided minimally invasive percutaneous treatment of spinal metastasis.

In order to provide effective options for minimally invasive treatment of spinal metastases, the present study retrospectively evaluated the efficacy and safety of image-guided minimally invasive percutaneous treatment of spinal metastases. Image-guided percutaneous vertebral body enhancement, radiofrequency ablation (RFA) and tumor debulking combined with other methods to strengthen the vertebrae were applied dependent on the indications. Percutaneous vertebroplasty (PVP) was used when vertebral body destruction was simple. In addition, RFA was used in cases where pure spinal epidural soft tissue mass or accessories (spinous process, vertebral plate and vertebral pedicle) were destroyed, but vertebral integrity and stability existed. Tumor debulking (also known as limited RFA) combined with vertebral augmentation were used in cases presenting destruction of the epidural soft tissue mass and accessories, and pathological vertebral fractures. A comprehensive assessment was performed through a standardized questionnaire and indicators including biomechanical stability of the spine, quality of life, neurological status and tumor progression status were assessed during the 6 weeks-6 months follow-up following surgery. After the most suitable treatment was used, the biomechanical stability of the spine was increased, the pain caused by spinal metastases within 6 weeks was significantly reduced, while the daily activities and quality of life were improved. The mean progression-free survival of tumors was 330±54 days, and no associated complications occurred. Therefore, the use of a combination of image-guided PVP, RFA and other methods is safe and effective for the treatment of spinal metastases.

Solvothermal synthesis of wire-like SnxSb2Te3+x with an enhanced thermoelectric performance.

Nanostructured tellurides have attracted increasing attention in thermoelectric applications for waste heat recovery and cooling devices. Here, we report on the synthesis of wire-like SnxSb2Te3+x (x = 0, 0.02 and 0.05) nanoparticles using elemental precursors in EG. The enhanced thermoelectric performance was achieved in alloyed samples due to the increase of carrier population in heavy valence band valleys by incorporating Sn(2+) at the Sb(3+) sublattice, enabling the simultaneous realization of low electrical resistivity along with a high Seebeck coefficient as well as the decline of thermal conductivity. Thus a boosted power factor and low thermal conductivity lead to the highest ZT value of 0.58 at 150 °C in the Sn0.02Sb2Te3.02 sample. Our research offers a general wet-chemical route for the preparation of one-dimensional nanomaterials and probably promotes the practical thermoelectric applications of Sb2Te3-based materials at low temperatures.

Allicin protects spinal cord neurons from glutamate-induced oxidative stress through regulating the heat shock protein 70/inducible nitric oxide synthase pathway.

Allicin, the main biologically active compound derived from garlic, exerts a broad spectrum of pharmacological activities and is considered to have therapeutic potential in many neurological disorders. Using an in vitro spinal cord injury model induced by glutamate treatment, we sought to investigate the neuroprotective effects of allicin in primary cultured spinal cord neurons. We found that allicin treatment significantly attenuated glutamate-induced lactate dehydrogenase (LDH) release, loss of cell viability and apoptotic neuronal death. This protection was associated with reduced oxidative stress, as evidenced by decreased reactive oxygen species (ROS) generation, reduced lipid peroxidation and preservation of antioxidant enzyme activities. The results of western blot analysis showed that allicin decreased the expression of inducible nitric oxide synthase (iNOS), but had no effects on the expression of neuronal NOS (nNOS) following glutamate exposure. Moreover, allicin treatment significantly increased the expression of heat shock protein 70 (HSP70) at both mRNA and protein levels. Knockdown of HSP70 by specific targeted small interfere RNA (siRNA) not only mitigated allicin-induced protective activity, but also partially nullified its effects on the regulation of iNOS. Collectively, these data demonstrate that allicin treatment may be an effective therapeutic strategy for spinal cord injury, and that the potential underlying mechanism involves HSP70/iNOS pathway-mediated inhibition of oxidative stress.

Curcumin attenuates Nrf2 signaling defect, oxidative stress in muscle and glucose intolerance in high fat diet-fed mice.

AIM: To investigate the signaling mechanism of anti-oxidative action by curcumin and its impact on glucose disposal. METHODS: Male C57BL/6J mice were fed with either a normal diet (n = 10) or a high fat diet (HFD) (n = 20) to induce obesity and insulin resistance. After 16 wk, 10 HFD-fed mice were further treated with daily curcumin oral gavage at the dose of 50 mg/kg body weight (BW) (HFD + curcumin group). After 15 d of the curcumin supplementation, an intraperitoneal glucose tolerance test was performed. Fasting blood samples were also collected for insulin and glucose measurements. Insulin-sensitive tissues, including muscle, adipose tissue and the liver, were isolated for the assessments of malondialdehyde (MDA), reactive oxygen species (ROS) and nuclear factor erythroid-2-related factor-2 (Nrf2) signaling. RESULTS: We show here that in a HFD mouse model, short-term curcumin gavage attenuated glucose intolerance without affecting HFD-induced BW gain. Curcumin also attenuated HFD-induced elevations of MDA and ROS in the skeletal muscle, particularly in its mitochondrial fraction, but it had no such an effect in either adipose tissue or the liver of HFD-fed mice. Correspondingly, in skeletal muscle, the levels of total or nuclear content of Nrf2, as well as its downstream target, heme oxygenase-1, were reduced by HFD-feeding. Curcumin intervention dramatically reversed these defects in Nrf2 signaling. Further analysis of the relationship of oxidative stress with glucose level by a regression analysis showed a positive and significant correlation between the area under the curve of a glucose tolerance test with MDA levels either in muscle or muscular mitochondria. CONCLUSION: These findings suggest that the short-term treatment of curcumin in HFD-fed mice effectively ameliorates muscular oxidative stress by activating Nrf2 function that is a novel mechanism for its effect in improving glucose intolerance.

[Clinical application of continuous douche and vacuum sealing drainage in refractory tissue, bone and joint infections after debridement].

OBJECTIVE: To explore effect and the application value of continuous douche and vacuum sealing drainage (VSD) in refractory tissue, and joint infections after complete debridement. METHODS: As retrospective analysis of treatment time and restoration or recurrence, from Jan. 2006 to Dec. 2007, 61 cases of refractory tissue, bone and joint infections underwent continuous douche and VSD combined with the treatment of anti-inflammatory and rehabilitation training after debridement in our hospital. The 61 patients included 39 males and 22 females with age ranging from 10 to 58 years with an average of (35 +/- 12) years, among whom 61 identified to have ankle ulcers combined with infections,open fracture combined with infections, sacrococcygeal pressure ulcers combined with infections, infections after hip replacement, infections after open fracture, and infections after skin avulsion postoperation were 11, 15, 9, 3, 5 and 18 cases respectively. The course was from 2 weeks to 11 months with an average of 4 months. RESULTS: In all 61 patients,the mean healing time was 17, 36, 42, 24, 32, 29 and 28 days in ankle ulcers and infections, tibia and fibula open fracture and infections, femoral shaft fracture and infections, sacrococcygeal pressure ulcers and infections, infections after hip replacement, infections after open fracture, and infections after skin avulsion postoperation respectively. The replacement of VSD was 1, 2-4, 3-5, 1-3, 2-4, 2-3 and 1-3 times in each group respectively. There was no wound recurrence except for 2 cases with recurrent in 61 cases with external fixation nail hole semi-pathological fracture in 1 case of femoral shaft fracture and infection and 1 case of tibia and fibula fracture and infection after follow-up at least one year. CONCLUSION: Application of continuous douche and VSD can effectively decrease incidence of complications and promote the refractory tissue, bone and joint infections wound growth, healing and considerably shorten the healing time.

[Environmental responses of four urban tree species transpiration in northern China].

By using thermal dissipation probes (TDP), this paper monitored the sap flow of four tree species (Cedrus deodara, Zelkova schneideriana, Euonymus bungeanus, and Metasequoia glyptostroboides) at the Laodong Park in Dalian City from June to August 2008, and the soil moisture content and micrometeorological variables were mehsured simultaneously. Due to the absence of water-stress in the habitat, the sap flow of all sampled trees had no significant correlation with soil moisture content (R2 < 0.050, P > 0.211, n=1296). The correlation coefficient between solar radiation and sap flow reached 0.624-0.773 (P = 0.00, n=1296) despite the existing hysteresis. Solar radiation had major effect (R2 > 0.700, P < 0.05) during early morning (5:00-8:00) and late afternoon (18:00-20:00) when undergoing dramatic changes. As the main factor determining nighttime sap flow (R2 > 0.660, P < 0.05, n=1872), vapor pressure deficit (VPD) had a correlation coefficient as high as 0.650-0.823 (P = 0.00, n=1296) with the sap flow in whole-day scale. Meanwhile, the models constructed on the basis of VPD were able to explain 90% of daily sap flow change (P = 0.00). The correlation coefficient between sap flow and wind speed was relatively smaller than the previous two (R2 < 0.380, P = 0.00, n=1296), though showing significant correlation in affecting sap flow. Observations also detected the saturation phenomenon of sap flow to the environmental demands.

[Reactive astrocytes and nestin expression in adult rats following spinal cord compression injury].

OBJECTIVE: To observe the expressions of nestin and glial fibrillary acidic protein (GFAP) and their association with reactive astrocytes following spinal cord injury in adult rats. METHODS: Adult rats with compression injury of the spinal cord were divided into 7 groups (n=6) and examined at 1, 3, and 5 days and at 1, 2, 4 and 8 weeks after the injury. The recovery of the locomotor function after the injury was evaluated with Basso, Beattie and Bresnahan (BBB) scale, and the degree and scope of the spinal injury were assessed using toluidine blue staining. Immunohistochemistry, double immunofluorescent labeling and an image analysis system were employed to observe nestin and GFAP expression and cell proliferation in different regions of the spinal cord. RESULTS: The bilateral hind limb locomotor function of the rats declined severely 24 h after the spinal cord injury and underwent substantial recovery in 1 or 2 weeks after the injury, but followed by rather slow recovery afterwards. Toluidine blue staining of the spinal cord 24 h after the injury showed significant pathological changes in the neurons. The extension of the tissue injury increased with time till 1 week after the spinal cord injury. The site of injury and the adjacent tissues presented with markedly increased nestin and GFAP expressions 24 h after the injury, and nestin+/GFAP(-) cells dominated in the ependymal region around the central canal, whereas nestin+/GFAP+ dominated in the in other regions, showing significant difference from the control group. Nestin and GFAP expression reached the peak level 3 to 7 days after the injury and declined gradually till reaching nearly the control level at 2 weeks. CONCLUSION: Compression injury of the spinal cord induces up-regulated expressions of nestin and GFAP, and nestin expression is positively correlated to the reactive astrocytes, which, along with the neural stem cells, respond to spinal nerve injury and possibly play a role in repair of the central nervous system injury.

[Clinical observation on effect of gufusheng capsule in treating early stage Legg-Calve-Perthes disease].

OBJECTIVE: To evaluate the clinical efficacy of Gufusheng capsule (GFSC) in treating early stage Legg-Calve-Perthes disease. METHODS: Adopting randomized single controlled trial, 45 cases with Legg-Calve-Perthes disease in early stage were randomly divided into 2 groups, 23 patients in the control group were treated with arthrectomy hip synovial membrane excision of affected lateral, femoral head decompression and transplantation of muscle-bone flap from ileum, above treatment were also given to the 22 patients in the treated group but combined with orally taking of GFSC. The treatment course for both groups was 6 months. RESULTS: Short-term effect after 6 months' treatment showed that the total effective rate in the treated group and the control group was 90.9% and 78.3% respectively, and the excellent rate was 72.7% and 60.9% respectively. The pain visual analogue scoring, clinical symptom and syndrome scoring markedly decreased before and after treatment, showing significant difference between the two groups (P < 0.05, P < 0.01). Long-term effect after 1-year to 4-year follow-up showed that the total effective rate and excellent rate between the treated group and the control group was significantly different ( chi2 = 8. 5976, P < 0.05). CONCLUSION: GFSC has marked effect in alleviating pain and ameliorating function of hip joint, being an effective compound recipe for treatment of early stage Legg-Calve-Perthes disease. Therefore, it is worthy of researching and developing furthermore.